Surrogate and Sentinal Technologies to Monitor Stability of Cancer Phosphoprotein

描述(由申请人提供)：磷酸化，或激酶驱动的信号网络的激活状态体现了关于疾病发病机制和与激酶相关的治疗靶点的持续状态的信息。使用人类肿瘤活检标本分析肿瘤磷酸蛋白质组是即将到来的个体化癌症治疗革命的关键组成部分。该应用程序尚未满足的一个关键需求是组织磷蛋白稳定性数据、标准化方案和可用于现实世界临床环境(例如，手术室、门诊活检、放射套件针吸检查)的新技术，用于磷蛋白的无缝采集、即时保存和实时稳定性监测。我们的多学科团队此前开发了激光捕获显微切割(LCM)和反相蛋白质微阵列(RPA)技术，用于对组织微环境进行磷酸蛋白质组分析。我们将标准化RPA技术，以高精度和高灵敏度同时测量至少100个已知的人体核心针吸活组织或细针抽吸物中的磷蛋白终点。这项技术将被用来收集以前未知的关于磷蛋白组织半衰期的定量信息，这些信息代表了一系列信号通路、细胞室和磷酸化残基在0到48小时的时间。这些数据将成为确定具有高度代表性和最不稳定的内源性磷蛋白的子集的基础，这些蛋白可以作为组织保存和磷蛋白稳定性的新的替代定量标记。我们将利用这些稳定性数据和新的替代终点技术，为组织易腐烂极限提出明确的定量指南。我们将创建外源磷蛋白前哨纳米颗粒来记录样品的加工历史，以进行持续的质量保证。RPA、替代标记和前哨技术将用于对在室温下稳定激酶、磷酸酶和磷蛋白24-48小时的候选防腐液进行评级。预期的结果将是一个完整的标准化技术工具包，可用于常规的临床收集、运输、对死亡组织的早期预警，以及用于分子特征分析的组织磷蛋白的实时监测。

DESCRIPTION (provided by applicant): The phosphorylation, or activation state of kinase-driven signal networks embodies information concerning disease pathogenesis and the ongoing state of kinase associated therapeutic targets. Profiling the tumor phosphoproteome using human tumor biopsy specimens is a crucial component of the perceived upcoming revolution of individualized cancer therapy. A critical unmet need addressed by this application is tissue phosphoprotein stability data, standardized protocols, and novel technologies which can be used in the real world clinical setting (e.g. operating room, outpatient clinic biopsy, radiological suite needle aspiration) for seamless collection, immediate preservation, and real time stability monitoring of phosphoproteins. Our multidisciplinary team has previously developed Laser Capture Microdissection (LCM) and reverse phase protein microarray (RPA) technologies to conduct phosphoproteomic analysis of the tissue microenvironment. We will standardize the RPA technology to simultaneously measure at least 100 known phosphoprotein endpoints with high precision and sensitivity in a human core needle biopsy or fine needle aspirate. This technology will be used to collect previously unknown quantitative information about the tissue half-life of phosphoproteins representing a wide range of signaling pathways, cellular compartments and phosphorylated residues over time zero to 48 hours. These data will become the basis to identify a subset of highly representative and most-labile endogenous phosphoproteins which can be employed as novel surrogate quantitative markers of tissue preservation and phosphoprotein stability. We will employ this stability data and novel surrogate endpoints technology to propose definitive quantitative guidelines for tissue perishability limits. We will create exogenous phosphoprotein sentinel nanoparticles to record the processing history of the specimen for ongoing Quality Assurance. The RPA, surrogate markers, and sentinel technology will be used to rank candidate preservative solutions that stabilize kinases, phosphatases, and phosphoproteins for 24-48 hr at room temperature. The desired outcome will be a complete standardized technology kit which can be employed for routine clinical collection, shipping, early warning of perished tissue, and real time monitoring of tissue phosphoproteins for molecular profiling.